银杏叶、瓜蒌、丹参及配伍制剂对兔心肌缺血再灌注损伤的保护作用

Protective Effect of Ginkgo Biloba,Trichosanthes Kirilowii Maxim,Salvia Miltiorrhiza Bunge and Their Compatibility on Myocardial Ischemia Reperfusion Injury Rabbits

目的以兔血液指标的波动和心肌组织病变为切入点,评价银杏叶丹参汤主要成分单体及配伍对心肌缺血再灌注损伤(MIRI)的保护作用。方法将新西兰兔50只随机分为模型对照组、银杏叶组、瓜蒌组、丹参组和配伍组,10只/组。各给药组灌胃给予相应药物,模型对照组给予等体积纯化水,持续1周。通过结扎兔冠脉左前降支30 min后再灌注120 min,制备心肌缺血再灌注损伤模型。检测造模前、缺血30 min和再灌注120 min兔血液一氧化氮(NO)、一氧化氮合成酶(NOS)、超氧化物歧化酶(SOD)、丙二醛(MDA)、白介素-6(IL-6)、肿瘤坏死因子α(TNF-α)和肌酸磷酸激酶同工酶(CK-MB)水平。HE染色观察心肌组织病理变化,凋亡细胞原位末端转移酶标记法(TUNEL)检测心肌细胞凋亡率。结果瓜蒌组MIRI 120 min血清NO含量、NOS活性高于模型对照组(P<0.05);配伍组缺血30 min、MIRI 120 min,银杏叶组MIRI 120 min血清NO含量均低于模型对照组(P<0.05);瓜蒌组心肌缺血30 min、MIRI 120 min,丹参组和配伍组MIRI 120 min血清SOD活性均高于模型对照组(P<0.05);银杏叶组MIRI 120 min及丹参组缺血30 min血清MDA含量均低于模型对照组(P<0.05);丹参组缺血30 min兔血清IL-6含量高于模型对照组(P<0.05);模型对照组MIRI 120 min血清IL-6含量低于丹参组,高于瓜蒌组(P<0.05);银杏叶组、瓜蒌组和配伍组MIRI 120 min血清TNF-α含量均高于模型对照组(P<0.05);银杏叶组、丹参组和配伍组缺血30 min,银杏叶组、瓜蒌组、丹参组和配伍组MIRI 120 min血清CK-MB含量低于模型对照组(P<0.05);银杏叶组、瓜蒌组、丹参组和配伍组MIRI兔心肌的凋亡率低于模型对照组,差异有统计学意义(P<0.05)。结论银杏叶、丹参、瓜蒌及其配伍制剂均可有效减轻MIRI兔心肌损伤,作用机制可能主要与抗氧化应激和调节炎症反应有关,配伍制剂作用机制较各单体提取物复杂。

Objective To study the protective effect of Ginkgo Biloba,Trichosanthes Kirilowii Maxim,Salvia miltiorrhiza Bunge and their compatibility on myocardial ischemia reperfusion injury(MIRI)of rabbits through observing changes of blood index and cardiac tissue.Methods A total of 50 conventional New Zealand rabbits were randomly divided into model control group,Ginkgo Biloba group,Trichosanthes Kirilowii Maxim group,Salvia Miltiorrhiza Bunge group and compatibility group,with 10 rabbits in each group.Each administration group was given corresponding drugs by gavage,and the model control group was given equal volume of purified water for 1 week.Myocardial ischemia-reperfusion injury model was prepared by ligating the left anterior descending coronary artery for 30 min and reperfusion for 120 min.The levels of nitric oxide(NO),nitric oxide synthase(NOS),superoxide dismutase(SOD),malondialdehyde(MDA),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and creatine kinase isoenzyme(CK-MB)in blood were detected before modeling,30 min after ischemia and 120 min after reperfusion.The pathological changes of myocardial tissue were observed by HE staining,and the apoptosis rate of myocardial cells was detected by TdT-mediated dUTP nick-end labeling(TUNEL).Results The serum NO content and NOS activity in the Trichosanthes Kirilowii Maxim group were higher than those in the model control group at 120 min after MIRI(P<0.05).The level of serum NO in the compatibility group at 30 min of ischemia,120 min of MIRI and 120 min of MIRI in the Ginkgo Biloba group was lower than those in the model control group(P<0.05).The serum SOD activity of myocardial ischemia 30 min,MIRI 120 min in the Trichosanthes Kirilowii Maxim group and MIRI 120 min in the Salvia Miltiorrhiza Bunge group and compatibility group was higher than that in the model control group(P<0.05).The level of serum MDA in Ginkgo Biloba group at 120 min of MIRI and Salvia Miltiorrhiza Bunge group at 30 min of ischemia was lower than those in model control group(P<0.05).The content of serum IL-6 in Salvia Miltiorrhiza Bunge group was higher than that in model control group(P<0.05).The content of serum IL-6 at 120 min of MIRI in the model control group was lower than that in the Salvia Miltiorrhiza Bunge group and higher than that in the Ginkgo Biloba group(P<0.05).The content of serum TNF-αat 120 min of MIRI in Ginkgo Biloba group,Trichosanthes Kirilowii Maxim group and compatibility group was higher than that in model control group(P<0.05).The levels of serum CK-MB in Ginkgo biloba group,Salvia Miltiorrhiza Bunge group and compatibility group at 30 min of ischemia,and Ginkgo biloba group,Trichosanthes Kirilowii Maxim group,Salvia Miltiorrhiza Bunge group and compatibility group at 120 min of MIRI was lower than that in model control group(P<0.05).The myocardial apoptosis rate of MIRI rabbits in Ginkgo biloba group,Trichosanthes Kirilowii Maxim group,Salvia Miltiorrhiza Bunge group and compatibility group was lower than that in model control group,and the difference was statistically significant(P<0.05).Conclusion Ginkgo Biloba,Trichosanthes Kirilowii Maxim,Salvia miltiorrhiza Bunge and their compatibility can reduce MIRI through regulating oxidative stress and inflammation,and mechanism of compatibility seems to be more complicated.